Gene expression of Paracoccidioides virulence factors after interaction with macrophages and fibroblasts

Abstract

BACKGROUND Paracoccidioidomycosis (PCM) is a systemic mycosis with high prevalence in Latin America that is caused by thermodimorphic fungal species of the Paracoccidioides genus.OBJECTIVES In this study, we used quantitative polymerase chain reaction (qPCR) to investigate the expression of genes related to the virulence of Paracoccidioides brasiliensis (Pb18) and P. lutzii (Pb01) strains in their mycelial (M) and yeast (Y) forms after contact with alveolar macrophages (AMJ2-C11 cell line) and fibroblasts (MRC-5 cell line).METHODS The selected genes were those coding for 43 kDa glycoprotein (gp43), enolase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 14-3-3 protein (30 kDa), phospholipase, and aspartyl protease. FINDINGS In the Pb18 M form, the aspartyl protease gene showed the highest expression among all genes tested, both before and after infection of host cells. In the Pb18 Y form after macrophage infection, the 14-3-3 gene showed the highest expression among all genes tested, followed by the phospholipase and gp43 genes, and their expression was 50-fold, 10-fold, and 6-fold higher, respectively, than that in the M form. After fibroblast infection with the Pb18 Y form, the 14-3-3 gene showed the highest expression, followed by the phospholipase and aspartyl protease genes, and their expression was 25-fold, 10-fold, and 10-fold higher, respectively, than that in the M form. Enolase and aspartyl protease genes were expressed upon infection of both cell lines. After macrophage infection with the Pb01 Y form, the 14-3-3 gene showed the highest expression, followed by the phospholipase and aspartyl protease genes, and their expression was 18-fold, 12.5-fold, and 6-fold higher, respectively, than that in the M form. MAIN CONCLUSIONSIn conclusion, the data show that the expression of the genes analysed may be upregulated upon fungushost interaction. Therefore, these genes may be involved in the pathogenesis of paracoccidioidomycosis.

Peptides derived from gp43, the most antigenic protein from Paracoccidioides brasiliensis, form amyloid fibrils in vitro: implications for vaccine development

“…The success of yeast-host interaction depends on several regulatory mechanisms as well as the expression of several different virulence factors. Among these are adhesins, which are surface proteins that recognize extracellular matrix (ECM) components of the host cells 2 , 3 .…”

Alternative Non-Mammalian Animal and Cellular Methods for the Study of Host–Fungal Interactions

“…Initially, it employed crude extracts and animal lymph as culture mediums. However, it has since evolved into an indispensable tool for investigating degenerative diseases, cell therapy, stem cell research, testing the toxicity and efficacy of novel compounds, and understanding pathogenesis, among other applications [ 16 , 17 ].…”

“…Despite its close resemblance to natural conditions, the in vitro process still presents challenges for cell development. Its cell–cell and cell–matrix adhesion is diminished, and it lacks the attributes (such as heterogeneity and three-dimensional architecture) of an in vivo tissue since its nutritional and hormonal environment undergo modification [ 17 , 24 ].…”

Alginate-Based 3D A549 Cell Culture Model to Study Paracoccidioides Infection

“…Current research in fungi–host interaction in vitro relies mainly on two-dimensional (2D) single-cell cultures. Although these methods have contributed to gathering knowledge about these interactions, infectious disease mechanisms, and the development of new therapeutic agents, they lack many essential features that are present in vivo and/or in animal models [ 1 , 2 , 3 ]. Furthermore, there is a lack of microenvironmental data regarding the respiratory tract’s main entry point for many fungi [ 4 , 5 , 6 ].…”